Sax C M, Farrell F X, Tobian J A, Zehner Z E
Department of Human Genetics, Massey Cancer Center, Richmond, VA 23298.
Nucleic Acids Res. 1988 Aug 25;16(16):8057-76. doi: 10.1093/nar/16.16.8057.
The expression of vimentin is unique within the intermediate filament multigene family. It is the only member which deviates from its usual tissue-specific expression pattern and whose 5'-flanking region contains multiple GC boxes, the binding site for Sp1. The activity of vimentin 5'-end:CAT fusions has been compared in cells where vimentin is highly expressed (mouse L cells) or not expressed at all (MH1C1). In addition, CAT activity has been examined by microinjection into Xenopus oocytes. Both in vivo expression and in vitro binding studies implicate Sp1 as a general regulatory factor in vimentin gene expression. Increased expression of 5'-end:CAT fusions in mouse L cells suggests that a fibroblast-specific enhancer element resides in the region -321 to -160. Low transcriptional activity in MH1C1 cells may be due to either the lack of this positive transcription factor(s) or the presence of a repressor element. Here, we demonstrate that the unique and complex pattern of vimentin gene expression is controlled by multiple cis-acting elements.
波形蛋白的表达在中间丝多基因家族中是独特的。它是唯一偏离其通常组织特异性表达模式的成员,其5'侧翼区域包含多个GC盒,即Sp1的结合位点。已在波形蛋白高表达的细胞(小鼠L细胞)或根本不表达的细胞(MH1C1)中比较了波形蛋白5'端:CAT融合体的活性。此外,通过显微注射到非洲爪蟾卵母细胞中检测了CAT活性。体内表达和体外结合研究均表明Sp1是波形蛋白基因表达中的一般调节因子。5'端:CAT融合体在小鼠L细胞中的表达增加表明成纤维细胞特异性增强子元件位于-321至-160区域。MH1C1细胞中低转录活性可能是由于缺乏这种正转录因子或存在阻遏元件。在这里,我们证明波形蛋白基因表达的独特而复杂的模式受多个顺式作用元件控制。